To Study The Prevention Of Lung Cancer Activities And Mechanisms Of Vitamin E On Passive Smoking Mice

ObjectiveThe influence of smoking to smokers has been known to all.However,hazard of cigarette smog to passive smokers isn't taken into account carefullyby people until the recent 10 years. It has been reported that 51.9 percent ofnon-smokers are suffering from passive smoking. Hazard of cigarette smog topassive smokers is undiscoverly ,extensively and seriously.Passive smokinghas become a concerned important enviromental hygiene problem.Smoke isthe main reason that results to lung cancer.Lung cancer is the highestincidence and mortality among all cancers.WHO suggests that a newapproaches for preventing cancers and anti-cancers is food-prevention.VE is acommon vitamin and it widely exits in human daily food. Epidemiologicalstudies have pointed that the food rich in VE may decrease the danger of theincidence of lung cancer. This thesis studies the prevention of lung canceractivities and mechanisms of VE on passive smoking KunMing mice byfounding passive smoking mice modle and the culture of ELC in vitro.We hopethe thesis can supports some valuable experimental datas for lung cancerprevention.Methods1 Experimental animals and groups150 healthy and clean KunMing mice (female 75, male 75) ,aged 5 weeks.Made mice into 3 groups:Normal control group(NC),somking controlgroup(SC) , smoking and VE group(SV).2 Toxins infection ways and VE intervening measureTook SC group's and SV group's mice into dynamic passive smokingmeasure,exposured for 6h a day,5 days a week for 5 months and a 4 monthsrecovery period in air and the NC group was kept in air during the entireperiod of 9 months.SE group mice received a dose of 100mg·kg-1·d-1 VE ondiet throughout the experiment. All animals were killed 9 months after thebeginning of the experiment.3 ELC culture in vitro and intervening measuresTook the 13th day embryo lung cells in RPMI-1640 with 10% FBS invitro culture.Producted CSE and measured nicotin concentration in it byHPLC. Different concentrations CSE affected ELC and differentconcentration VE intervened.4 Figures and Methods4.1 Lung cancer incedence of passvie smoking mice measures: Number of lungtumors;pathologic diagnose of lung tumors (HE dye)4.2 Body weight change and mortality of passive smoking mice4.3 Effect of VE of immunity ability on passive smoking mice : thymusindex;spleen index;T Lymphocyte Transformation Test4.4 Effect of VE on anti-oxidant activities on passive smoking mice: ROS;VE;T-SOD;MDA;GSH-Px4.5 Effect of VE of lung cell on passive smoking mice:Cell cycle and Apoptosisrate (FCM)4.6 Effect of CSE of on ELC:Prolifertion (MTT assay);Cell cycle andApoptosis rate(FCM)4.7 Effect of CSE and VE of on ELC:Prolifertion (MTT assay);Cell cycle andApoptosis rate(FCM)Result1 The effect of VE on incedence rate and multiplicity of lung tumor in passivesmoking mice.Passive smoking resulted to lung tumors incedence and lungtumors multiplicity.The SC group's were higer than NC group's(p<0.05),but the SV group's were lower than the SC group'(sp<0.05). However,therehad no obvious difference between the SV group's and the NC group's(p>0.05).2 The effect of VE on the body weight change and the mortality in passivesmoking miceIn the five passive smoking months,the body weight increase of smokinggroups (SC and SV)mice were slower than NC group's.Then,the bodyweight increasing of SV group mice was faster than the other two groups' inthe four-month recovery times.At the end of experiment,the body weight ofSC group was lower than the other two groups' (p<0.05). However,therehad no obvious difference between the SV group's and the NC group's(p>0.05).The mortality of NC groupwas obviously higher than the othertwo groups' (p<0.05),but there had no obvious difference between the SVgroup's and the NC group's(p>0.05).3 The effect of VE on immunity ability in passive smoking miceThe results of lyphocyte transformation test showed that passive smokingdereased SC group mice's SI(p<0.05).However,VE increased SI of passivesmoking mice(p<0.05).The spleen index of SC group mice was lower thanthe other two groups(p<0.05),but there there had no obvious differencebetween the SV group's and the NC group's(p>0.05).And there had nodifference among the thymus index of three groups(p>0.05).4 The effect of VE on anti-oxidant activities in passive smoking micePassive smoking producted a plenty of ROS in mice serum.VE obviouslydecreased ROS(SV vs SC,p<0.01).Furthermore, VE obviously decreasedMDA concentration of passive smoking mic(eSV vs SC,p<0.05).Besides, VEobviously strengthened T-SOD and GSH-PX activities of passive smokingmice(SV vs SC,p<0.01).5 The effect of VE on the cell cycle and apoptosis rate of lung cell in passivesmoking micePassive smoking made lung cell of passive smoking mice apoptosi(sSC vsNC,p<0.05).But VE had no influence on apoptosis and cell cycle of lung cellin passive smoking mice(SV vs SC,p>0.05).6 The effect of CSE on ELC in vitroThe results in vitro showed that CSE inhibited ELC perliferation.Theinhibitation had time-dependent relation and dose-dependentrelation.Especially, after 24h,48h and 72h of exposure to CSE ,inhibition ofELC proliferation was observed at high ECS of nicotine concentration 1:350or greater. Complete inhibition of cellular proliferation was observed atconcentrations of 1:1.The effect of CSE on cell cycle of ELC showed thatCSE made ELC prevented G0/G1 period to S period(p<0.05).CSE made ELCapoptosis. Especially, CSE of nicotine in high concentration (1:350, 1:175,1:1) (vs control,p<0.01).Meanwhile,light moricgraphs proved that CSEmade ELC fall off.7 The effect of VE and CSE on ELC in vitroThe results showed that VE strenghtened ELC perliferation that CSEtreated.The effect had time-dependent relation and dose-dependentrelation.Especially, VE of 100μmol·L-1 obviously inhibited ELC(vs CSEcontrol ,p<0.05).The effect of VE on cell cycle and apoptosis of ELC treatedby CSE showed that VE prevented ELC from CSE demage .Meanwhile,lightmoricgraph proved that VE obviously protected ELC.Conclusion1 VE had the prevention activities lung cancer in passive smoking mice.VEobviously decreased the incedence and lung tumors multiplicity.2 VE weakened the influence that passive smoking reduced body weight andobviously decreased the mortality of passive smoking mice.3 VE strengthened immunity ability and anti-oxidant activities of passivesmoking mice.4 There had no influence on lung cell apoptosis and cell cycle in passivesmoking mice.5 VE prevented CSE to damage ELC...